Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis
UKMYC5 is a 96-well microtitre plate designed by the Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) to enable the measurement of minimum inhibitory concentrations (MICs) of 14 different anti-TB compounds for >30,000 clinical tuberculosis isolates....
| Huvudupphovsmän: | , , , , , , , , , , , |
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| Materialtyp: | Journal article |
| Språk: | English |
| Publicerad: |
American Society for Microbiology
2018
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| _version_ | 1826304776336310272 |
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| author | Rancoita, PMV Cugnata, F Cruz, AL Borroni, E Hoosdally, SJ Walker, TM Grazian, C Davies, TJ Peto, TEA Crook, DW Fowler, PW Cirillo, DM |
| author_facet | Rancoita, PMV Cugnata, F Cruz, AL Borroni, E Hoosdally, SJ Walker, TM Grazian, C Davies, TJ Peto, TEA Crook, DW Fowler, PW Cirillo, DM |
| author_sort | Rancoita, PMV |
| collection | OXFORD |
| description | UKMYC5 is a 96-well microtitre plate designed by the Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) to enable the measurement of minimum inhibitory concentrations (MICs) of 14 different anti-TB compounds for >30,000 clinical tuberculosis isolates. Unlike the MYCOTB plate, on which UKMYC5 is based, the plate included two new (bedaquiline and delamanid) and two repurposed (clofazimine and linezolid) compounds. UKMYC5 plates were tested by seven laboratories on four continents using a panel of 19 external quality assessment (EQA) strains, including H37Rv. To assess the optimal combination of reading method and incubation time, MICs were measured from each plate by two readers using three methods (mirrored-box, microscope and Vizion™ Digital viewing system) after 7, 10, 14 and 21 days incubation. In addition, all EQA strains were whole-genome sequenced and phenotypically characterized by 7H10/7H11 agar proportion method (APM) and MGIT960. We conclude that the UKMYC5 plate is optimally read using the Vizion™ system after 14 days incubation, achieving an inter-reader agreement of 97.9% and intra- and inter-laboratory reproducibility of 95.6% and 93.1%, respectively. The mirrored-box had similar reproducibility. Strains classified as resistant by APM, MGIT960 or the presence of mutations known to confer resistance consistently record elevated MICs compared with those strains classified as susceptible. Finally, the UKMYC5 plate records intermediate MICs for one strain which the APM measured MICs close the applied critical concentration, providing early evidence that the UKMYC5 plate can quantitatively measure the magnitude of resistance to anti-TB compounds due to specific genetic variation. |
| first_indexed | 2024-03-07T06:22:54Z |
| format | Journal article |
| id | oxford-uuid:f3472655-3545-446e-acac-e44cc15d5509 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T06:22:54Z |
| publishDate | 2018 |
| publisher | American Society for Microbiology |
| record_format | dspace |
| spelling | oxford-uuid:f3472655-3545-446e-acac-e44cc15d55092022-03-27T12:10:58ZValidating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosisJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:f3472655-3545-446e-acac-e44cc15d5509EnglishSymplectic Elements at OxfordAmerican Society for Microbiology2018Rancoita, PMVCugnata, FCruz, ALBorroni, EHoosdally, SJWalker, TMGrazian, CDavies, TJPeto, TEACrook, DWFowler, PWCirillo, DMUKMYC5 is a 96-well microtitre plate designed by the Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) to enable the measurement of minimum inhibitory concentrations (MICs) of 14 different anti-TB compounds for >30,000 clinical tuberculosis isolates. Unlike the MYCOTB plate, on which UKMYC5 is based, the plate included two new (bedaquiline and delamanid) and two repurposed (clofazimine and linezolid) compounds. UKMYC5 plates were tested by seven laboratories on four continents using a panel of 19 external quality assessment (EQA) strains, including H37Rv. To assess the optimal combination of reading method and incubation time, MICs were measured from each plate by two readers using three methods (mirrored-box, microscope and Vizion™ Digital viewing system) after 7, 10, 14 and 21 days incubation. In addition, all EQA strains were whole-genome sequenced and phenotypically characterized by 7H10/7H11 agar proportion method (APM) and MGIT960. We conclude that the UKMYC5 plate is optimally read using the Vizion™ system after 14 days incubation, achieving an inter-reader agreement of 97.9% and intra- and inter-laboratory reproducibility of 95.6% and 93.1%, respectively. The mirrored-box had similar reproducibility. Strains classified as resistant by APM, MGIT960 or the presence of mutations known to confer resistance consistently record elevated MICs compared with those strains classified as susceptible. Finally, the UKMYC5 plate records intermediate MICs for one strain which the APM measured MICs close the applied critical concentration, providing early evidence that the UKMYC5 plate can quantitatively measure the magnitude of resistance to anti-TB compounds due to specific genetic variation. |
| spellingShingle | Rancoita, PMV Cugnata, F Cruz, AL Borroni, E Hoosdally, SJ Walker, TM Grazian, C Davies, TJ Peto, TEA Crook, DW Fowler, PW Cirillo, DM Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title | Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title_full | Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title_fullStr | Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title_full_unstemmed | Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title_short | Validating a 14-drug microtitre plate containing bedaquiline and delamanid for large-scale research susceptibility testing of Mycobacterium tuberculosis |
| title_sort | validating a 14 drug microtitre plate containing bedaquiline and delamanid for large scale research susceptibility testing of mycobacterium tuberculosis |
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